(Revised Abstract) DESCRIPTION (provided by applicant): The overall hypothesis to be addressed in Project 2 is that a mild, fever-like increase in systemic temperature serves as an adjuvant to the immune response, and in combination with immunotherapy, can provoke more effective anti-tumor- immunity. This hypothesis is consistent with the theory that a physiological increase in temperature might serve as a systemic 'danger signal' to the immune system and is supported by substantial preliminary and published data from this group and work of others indicating that there are several thermally sensitive events in the innate and adaptive immune response. These events are very likely the basis for the anti-tumor effects noted earlier by this group when animals are treated with a fever-like whole body hyperthermia protocol. Project 2 focuses on whether the efficacy of vaccines or adoptive T cell immunotherapies in animal models can be improved by addition of heat. Aim 1 is to more fully define the ability of heat to enhance the immunogenicity of 2 clinically important cancer vaccine antigens (Her2/neu and gp 100) and, as described by Dr. Subjeck in Project 1, those same antigens complexed to HSP 110 and Grp 170. Several basic parameters that define adjuvancy will be evaluated. Aim 2 is to evaluate the ability of heat to enhance the anti-tumor potential of these same antigens, using several different tumor models. In each of these aims we will be dissecting the role of CD8+, CD4+ T cells and antibody production in the heat-regulated, anti-tumor immune response. We will also evaluate the development of long-term memory responses and the control of minimal residual disease settings. Aim 3 is to evaluate the ability of heat to act as an adjuvant in an adoptive immunotherapy setting with tumor antigen-specific cytolytic T cells. In keeping with the overall integration and interdependency of the Program, the antigens to be evaluated in Aims 1 and 2 are the same as those studied and developed by Dr. Subjeck in Project 1. And, in all three aims, we will be interacting closely with Dr. Evans in Project 3, correlating the immunological and anti-tumor effects seen in each combination therapy with the recruitment and tumor-homing potential of immune effector cells. As a result, the experimental design and execution of these experiments will benefit from extensive use of the Immunomonitoring and Thermal Treatment Core and the Biometrics Core. We will also be continuously interacting with our clinical team and with Dr. Greco of the Biometrics Core for identification of additional preclinical information needed for optimization of new trials as the immunological data and effects on tumor growth are collected. This is the overall, long-term goal of this Project.